Clinical Chemistry Exam 1 Lecture 6 – Flashcards

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True or False:
Enzymes are found in All body tissues
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True
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Many enzymes are present in ___ from normal cellular turnover
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Plasma
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Enzymes are ____ ____ and _____, not altered or consumed during the reaction
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Biologic proteins
Catalysts
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_____ amounts of Enzyme are released following cellular damage
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Increased
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Basic Enzyme Reaction
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E + S ES E + P
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Substrate Concentration is what order kinetics?
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First
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First Order Kinetics is directly proportional to what?
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Substrate concentration
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In First Order Kinetics, what is provided in excess?
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Enzyme
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At some point in First Order Kinetics, substrate will be consumed and ____ ____ is reached
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Maximum Velocity
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Michaelis-Menton Constant (Km)
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Mathematical expression of the relationship between velocity and substrate
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M-Mm Constant (Km) =
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Substrate concentration at which the enzyme yields half the possible maximum velocity
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Which two types of plots provide visualization of first order kinetics?
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M-M and Lineweaver-Burk
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Enzyme Concentration is what order Kinetics?
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Zero
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What is in excess during Zero Order Kinetics?
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Substrate
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True or False:
With Zero Order Kinetics, the Reaction rate depends on enzyme concentration
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True
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With Zero Order Kinetics what is supplied by the patient?
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Enzyme
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With Zero Order Kinetics what is supplied by manufactured reagent?
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Substrate
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Why is it called Zero Order Kinetics?
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When the rate of velocity no longer increase, velocity rates level off, signalling "zero" amount of increase in rate
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pH
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Extremes will denature or change ionic state of enzymes
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Most enzymes function at what pH?
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7-8
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pH is controlled in labs by what?
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Buffer solutions
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Increasing the temperature increases the ____ ____ to a point
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Reaction Rate
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For every ___ degree rise, the rate doubles until the enzyme is denatured
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10
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At what temperature will the enzyme usually denature?
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>40 degrees Celsius
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Most assays are run at what temperatures?
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25, 30, 37 degrees Celsius
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True or False:
Avoid repeated freeze-thaw
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True
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If testing is delayed what should you do with the sample to preserve the enzyme levels?
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Freeze or Refrigerate
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True or False:
It takes a few minutes for ALL enzymes to combine with substrate
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True
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The reaction time is what phase of the reaction?
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Linear
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About ______ or more minutes into testing, the reaction rate begins to diminish
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6
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Measurements should be taken during what phase of the reaction?
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Linear
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Cofactors
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Nonprotein molecules that may be necessary for proper enzyme function
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Cofactor Activators
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Inorganic molecules
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Name 3 Cofactor Activators
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Calcium
Magnesium
Other Metallic ions
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Cofactor Coenzymes
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Organic Molecules
Often provided in the testing reagent
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Name 2 Cofactor Coenzymes
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NAD
Vitamin B family
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What is the following an example of?
Creatine + ATP --(CK / Mg)--> Creatine Phosphate + ADP
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Activators
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What is the following an example of?
Pyruvate + NADH --(LD)--> Lactate + NAD
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Coenzymes
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What are 7 Factors that influence Enzyme Reactions?
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Substrate Concentration
Enzyme Concentration
pH
Temperature
Reaction Time
Cofactors
Inhibitors
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Competitive Inhibitors
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Bind to the active site of an enzyme
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What do you add to overcome Competitive Inhibitors?
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Substrate
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Is Competitive Inhibitors possible to anticipate?
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No
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Noncompetitive Inhibitors
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Bind to enzyme at a place other than the active site
Causes structural changes to the enzyme
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True or False:
Adding more substrate may not help with noncompetitive inhibitors
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True
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True or False:
The results of Noncompetitive inhibitors may be irreversible
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True
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Assays measure enzyme _____ which is a reflection of concentration
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activity
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What can be used to measure enzyme activity?
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Increase in product
Decrease in substrate
Decrease in coenzyme
Increase of an altered coenzyme
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In what Order are Measurements of Enzyme Activity always performed?
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Zero
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What must be in excess when measuring enzyme activity?
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Coenzyme
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Fixed-Time Method (End Point)
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The reaction proceeds for a designated time and is then stopped
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The velocity of a Fixed-Time Method is assumed to be what?
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Linear and proportional to enzyme concentration
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Continuous-Monitoring Method (Kinetic Analysis)
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Multiple measurements of absorbance are made every 30-60 seconds OR continuously
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True or False:
Continuous-Monitoring Method - Linearity may be more adequately verified
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True
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A sudden _____ in the reaction rate can be readily observed in the Continuous-Monitoring Method
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Decrease
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If a change in Absorption is not constant, then an ____ _____ is present
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Unknown Variable
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If the change in Absorption is not constant it may result from what?
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high levels of patient enzymes rapidly depleting the substrate
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What should you do if the change is absorption is not constant?
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Dilute patient sample and re-run
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International Unit (U or IU)
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The amount of enzyme that will catalyze the reaction of 1 micromole of substrate per minute under specified conditions of temperature, pH, substrates, and activators
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The International Unit is in what concentration?
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Units per Liter (U/L or IU/L)
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If an enzyme assay is 150 IU/L at 25C, what will be the product concentration if the assay is run at 35C?
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300 IU/L
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